Nadph oxidase inhibitors, pharmaceutical composition comprising the same, and application thereof

ABSTRACT

The present disclosure relates to a compound of Formula I, or a geometric isomer, enantiomer, diastereomer, racemate, atropisomer, pharmaceutically acceptable salt, prodrug or solvate thereof. The present disclosure further relates to a composition comprising the compound of Formula (I). The compound and the composition described herein can be used to inhibit NADPH oxidase activity.

BACKGROUND

Reactive Oxygen Species (ROS) are a number of reactive molecules andfree radicals derived from molecular oxygen, including superoxide anion(.O₂ ⁻), peroxide (.O₂ ⁻²), hydrogen peroxide (H₂O₂), hydroxyl radical(.OH) and hydroxyl radical ion (OH⁻). These molecules may be produced asbyproducts during the aerobic respiration or by oxidoreductase enzymesand metal catalyzed oxidation. Normally, ROS play a role in immunity andcell signaling, including apoptosis, gene expression, and the activationof cell signaling cascades. However, excessive ROS production and/orimpaired ROS reduction may lead to overload of ROS and oxidative stress.

Under conditions of oxidative stress, an imbalance between the systemicmanifestation of ROS and biological system's ability to neutralize thereactive intermediates or to repair the resulting damage is presented,the result of which was demonstrated to contribute to various acute andchronic diseases related to cardiovascular, inflammatory, oncological,and neurological therapeutic areas.

Several enzymes found throughout the cell are involved in ROSgeneration, including NADPH oxidase (NOX), xanthine oxidase,lipoxygenase, cyclooxygenases (COX) and substrate coupled nitric oxidesynthetase. Among the enzymes, NOX may produce a large amount of ROS innonphagocytic cells in both normal and pathological conditions, which isparticularly worthy of attention; because by inhibiting NOX activity,one may ameliorate oxidative stress without compromising immunity of thephagocytic cells.

SUMMARY

The disclosure relates to a series of NADPH oxidase inhibitors, apharmaceutical composition comprising the same, and a method ofinhibiting NADPH oxidase (NOX) activity.

According to one embodiment, a compound according to Formula (I) isprovided. The pharmaceutically acceptable salt, hydrate or solvate ofthis compound are also provided.

wherein A is C₆-C₂₀ aryl optionally fused with C₁-C₂₀ heterocyclyl, orC₁-C₂₀ heteroaryl optionally fused with C₁-C₂₀ heterocyclyl, and A isoptionally substituted with one or more C₁-C₁₀ alkyl, C₆-C₂₀ aryl,halogen, OR₁₁, C(O)R₁₁, C(O)OR₁₁, NR₁₁R₁₂, SO₂R₁₁, or SO₂(OR₁₁); B isC₁-C₁₀ alkyl, C₆-C₂₀ aryl, C₁-C₂₀ heteroaryl or C₁-C₂₀ heterocyclyl andB is optionally substituted with one or more C₁-C₁₀ alkyl, C₆-C₂₀ aryl,C₁-C₂₀ heterocyclyl, halogen, OR₂₁, C(O)R₂₁, C(O)OR₂₁, NR₂₁R₂₂, SO₂R₂₁,or SO₂(OR₂₁), wherein said C₁-C₁₀ alkyl is optionally substituted withone or more halogen; R₁₁, R₁₂, R₂₁ and R₂₂ independently are H, O,C₁-C₁₀ alkyl, or C₆-C₂₀ aryl, wherein said C₁-C₁₀ alkyl is optionallysubstituted with one or more halogen;

is a single bond or double bond; n is 0 or 1; X is N or C; Y is H, OH orNH₂; Z is nil or C(O); R₃ and R₄ independently are nil, H, C₁-C₁₀ alkylor C₆-C₂₀ aryl; and R₅ is nil, H or C₁-C₁₀ alkyl.

According to other embodiment, a pharmaceutical composition is provided.The pharmaceutical composition comprises the compound according toFormula I and at least a pharmaceutically acceptable excipient orcarrier.

According to still other embodiment, a method of inhibiting NADPHoxidase (NOX) is provided. The method comprises the step ofadministering to a cell an effective amount of the compound according toFormula I or the pharmaceutical composition comprising the compoundaccording to Formula I and at least a pharmaceutically acceptableexcipient or carrier.

DETAILED DESCRIPTION

The foregoing and other aspects of the present disclosure will now bedescribed in more detail with respect to other embodiments describedherein. It should be appreciated that the invention can be embodied indifferent forms and should not be construed as limited to theembodiments set forth herein. Rather, these embodiments are provided sothat this disclosure will be thorough and complete, and will fullyconvey the scope of the invention to those skilled in the art.

The terminology used in the description of the invention herein is forthe purpose of describing particular embodiments only and is notintended to be limiting of the invention. As used in the description ofthe invention and the appended claims, the singular forms “a”, “an” and“the” are intended to include the plural forms as well, unless thecontext clearly indicates otherwise.

As used herein, the terms “comprises,” “comprising,” “includes,”“including,” “has,” “having,” “contains”, “containing,” “characterizedby” or any other variation thereof, are intended to cover anon-exclusive inclusion, subject to any limitation explicitly indicated.For example, a composition, mixture, process or method that comprises alist of elements is not necessarily limited to only those elements butmay include other elements not expressly listed or inherent to suchcomposition, mixture, process, or method.

The transitional phrase “consisting of” excludes any element, step, oringredient not specified. If in the claim, such would close the claim tothe inclusion of materials other than those recited except forimpurities ordinarily associated therewith. When the phrase “consistingof” appears in a clause of the body of a claim, rather than immediatelyfollowing the preamble, it limits only the element set forth in thatclause; other elements are not excluded from the claim as a whole.

All of the carbon, hydrogen, oxygen, Sulfur, halogen, or nitrogeninvolved in the groups and compounds according to the present disclosureare optionally further replaced by one or more of their correspondingisotopes, wherein the carbon isotopes include ¹²C, ¹³C and ¹⁴C, thehydrogen isotopes include protium (H), deuterium (D, also known as heavyhydrogen) and tritium (T, also known as superheavy hydrogen), the oxygenisotopes include ¹⁶O, ¹⁷O and ¹⁸O, the sulfur isotopes include ³²S, ³³S,³⁴S and ³⁶S, the nitrogen isotopes include ¹⁴N and ¹⁵N, the fluorineisotopes include ¹⁷F and ¹⁹F, the chlorine isotopes include ³⁵Cl and³⁷Cl, and the bromine isotopes include ⁷⁹Br and ⁸¹Br.

“Alkyl” means a linear or branched saturated aliphatic hydrocarbylhaving 1 to 20 carbon atoms, preferably 1 to 10 carbon atoms.Non-limiting examples thereof include methyl, ethyl, n-propyl,isopropyl. n-butyl, sec-butyl, t-butyl, isobutyl, n-pentyl, isopentyl,neo-pentyl, n-hexyl, octyl, and various branched isomers thereof.

“Alkenyl” means a linear or branched unsaturated aliphatic hydrocarbylhaving 1 to 3 carbon-carbon double bonds, and comprising 2 to 20 carbonatoms, preferably 2 to 8. Non limiting examples thereof include vinyl,propen-2-yl, buten2-yl, buten-2-yl, penten-2-yl, penten-4-yl,hexen-2-yl, hexen3-yl, hepten-2-yl, hepten-3-yl, hepten-4-yl,octen-3-yl, nonen-3-yl, decen-4-yl and hendecen-3-yl. “Alkenyl” can alsoinclude polyenes such as 1,2 propadienyl and 2,4 hexadienyl.

“Alkynyl” means a linear or branched unsaturated aliphatic hydrocarbylhaving 1 to 3 carbon-carbon triple bonds, and comprising 2 to 20 carbonatoms, preferably 2 to 8 carbon atoms. Non limiting examples thereofinclude ethynyl, propyn-1-yl, propyn-2-yl, butyn-1-yl, butyn-2-yl,butyn-3-yl, 3.3-dimethylbutyn-2-yl, pentyn-1-yl, pentyn-2-yl,hexyn-1-yl, heptyn-1-yl, heptyn-3-yl, heptyn-4-yl, octyn-3-yl,nonyn-3-yl, decyn4-yl, hendecyn-3-yl or dodecyn-4-yl. “Alkynyl” can alsoinclude moieties comprised of multiple triple bonds such as 2,5hexadiynyl.

“Alkoxy” means —O-alkyl. Non-limiting examples thereof include methoxy,ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, t-butoxy,n-pentyloxy, n-hexyloxy, cyclopropoxy, and cyclobutoxy. This definitionapplies to the alkoxys used throughout this Description.

The term “halogen”, either alone or in compound words such as“haloalkyl”, or when used in descriptions such as “alkyl substitutedwith halogen” includes fluorine, chlorine, bromine or iodine. Further,when used in compound words such as “haloalkyl”, or when used indescriptions such as “alkyl substituted with halogen” said alkyl may bepartially or fully substituted with halogen atoms which may be the sameor different. Non-limiting examples of “haloalkyl” or “alkyl substitutedwith halogen” include F₃C—, ClCH₂—, CF₃CH₂— and CF₃CCl₂—.

The chemical abbreviations S(O) and S(═O) as used herein represent asulfinyl moiety. The chemical abbreviations SO₂, S(O)₂ and S(═O)₂ asused herein represent a sulfonyl moiety. The chemical abbreviations C(O)and C(═O) as used herein represent a carbonyl moiety. The chemicalabbreviations CO₂, C(O)O and C(═O)O as used herein represent anoxycarbonyl moiety. “CHO” means formyl.

“Aromatic” indicates that each of the ring atoms is essentially in thesame plane and has a p-orbital perpendicular to the ring plane, and that(4n+2) 7! electrons, where n is a positive integer, are associated withthe ring to comply with Hickel's rule. The term “aromatic ring or ringsystem” denotes a carbocyclic or heterocyclic ring or ring system inwhich the ring or at least one ring of the ring system is aromatic. Theterm “aromatic ring or ring system” is also referred to as “aryl”.“Aryl” might include 6 to 20 carbons as ring member including phenyl,benzyl, naphthyl, and the like. Aryl can also include substituted arylgroups such as tolyl. The term “aromatic heterocyclic ring system”denotes a heterocyclic ring system in which at least one ring of thering system is aromatic. The term “aromatic heterocyclic ring system” isalso referred to as “heteroaryl”. “Heteroaryl” might include 1 to 20carbons as ring member and might further include at least one heteroatomselected from S, N, and 0.

“Carbocyclyl” means a saturated or unsaturated aromatic or non-aromaticring, and the aromatic or non-aromatic ring may be a 3- to 8-memberedmonocyclic, a 4- to 12-membered bicyclic or a 10- to 20-memberedtricyclic system. The carbocyclic group may have attached bridge ringsor spiral rings. Non-limiting examples thereof include cyclopropyl,cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclodecyland cyclododecyl, cyclohexenyl,

This definition applies to the carbocyclyls used throughout thisDescription.

“Heterocyclyl” means a substituted or unsubstituted, saturated orunsaturated, aromatic or non-aromatic ring, and the aromatic ornon-aromatic ring may be a 1 to 8-membered monocyclic, a 4- to12-membered bicyclic or a 10- to 20-membered tricyclic system, andcontains at least one heteroatom selected from N, O or S. The optionallysubstituted N or S on the ring of a heterocyclic group may be oxidizedinto various oxidation States. The carbocyclic group may be attached viaa heteroatom or a carbon atom, and may have attached bridge rings orspiral rings. Non-limiting examples thereof include epoxyethyl,azacyclopropyl, oxzcyclobutyl, azacyclobutyl, 1,3-dioxolane,1,4-dioxolane, 1,3-dioxane, azacycloheptyl, pyridinyl, furyl,thiophenyl, pyranyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl,pyridazinyl, imidazolyl, piperidinyl, hexahydropyridinyl, morpholinyl,thiomorpholinyl, 1,3-dithia-, dihydrofuryl, dihydropyranyl,dithiacyclopentyl, tetrahydrofuryl, tetrahydropyrrolyl,tetrahydroimidazolyl, tetrahydrothiazolyl, tetrahydropyranyl,benzimidazolyl, benzopyridinyl, pyrrolopyridinyl, benzodihydrofuryl,azabicyclo[3.2.1]octyl, azabicyclo[5.2.0]nonyl,azatricyclo[5.3.1.1]dodecyl, aza-adamantanyl, and oxaspiro[3.3]heptyl.This definition applies to the heterocyclyls used throughout thisDescription.

As used herein, the term “optional” or “optionally” means the event orsituation modified by this term may but does not certainly happen,including both the case where the event or situation happens and thecase not. As used herein, the term “optionally substituted” refers togroups which are unsubstituted or have at least one non-hydrogensubstituent that does not extinguish the biological activity possessedby the unsubstituted analog. As used herein, the following definitionsshall apply unless otherwise indicated. The term “optionallysubstituted” is used interchangeably with the phrase “substituted orunsubstituted” or with the term “(un)substituted.” Unless otherwiseindicated, an optionally substituted group may have a substituent ateach substitutable position of the group, and each substitution isindependent of the other. For example, “a heterocyclyl optionallysubstituted with alkyl” means that the alkyl may be present but is notnecessarily present, including both the case where the heterocyclyl issubstituted with alkyl and the case where the heterocyclyl is notsubstituted with alkyl.

As used herein, a “pharmaceutically acceptable” component (such as acarrier or excipient) means that the compound or composition is suitablefor administration to a subject to achieve the treatments describedherein, without unduly deleterious side effects in light of the severityof the disease and necessity of the treatment. “Carrier” means amaterial that does not cause significant stimulation to an organism anddoes not eliminate the biological activity and characteristics of agiven compound. “Excipient” means an inert substance added into apharmaceutical composition to facilitate administration of a compound.Non-limiting examples thereof include calcium carbonate, calciumphosphate, sugar, starch, cellulose derivatives (includingmicrocrystalline cellulose), gelatin, vegetable oils, polyethyleneglycols, diluent, a granulating agent, lubricant, binder anddisintegrant.

As used herein, the term “effective amount” refers to the amount of eachactive agent required to confer the desired effect (e.g. inhibitingNADPH oxidase) on the subject, either alone or in combination with oneor more other active agents. An effective amount varies, as recognizedby those skilled in the art, depending on the particular condition beingtreated, the severity of the condition, the individual patientparameters including age, physical condition, size, gender and weight,the duration of the treatment, the nature of concurrent therapy (ifany), the specific route of administration and like factors within theknowledge and expertise of the health practitioner. These factors arewell known to those of ordinary skill in the art and can be addressedwith no more than routine experimentation. It is generally preferredthat a maximum dose of the individual components or combinations thereofbe used, that is, the highest safe dose according to sound medicaljudgment. It will be understood by those of ordinary skill in the art,however, that a patient may insist upon a lower dose or tolerable dosefor medical reasons, psychological reasons or for virtually any otherreasons.

A “stereoisomer” refers to an isomer of a molecule having its atoms in adifferent spatial arrangement, including cis-trans-isomer, enantiomer,and conformer. Compounds of this invention can exist as one or morestereoisomers. The various stereoisomers include enantiomers,diastereomers, racemates, atropisomers and geometric isomers.Stereoisomers are isomers of identical constitution but differing in thearrangement of their atoms in space and include enantiomers,diastereomers, cis-trans isomers (also known as geometric isomers) andatropisomers. Atropisomers result from restricted rotation about singlebonds where the rotational barrier is high enough to permit isolation ofthe isomeric species. One skilled in the art will appreciate that onestereoisomer may be more active and/or may exhibit beneficial effectswhen enriched relative to the other stereoisomer(s) or when separatedfrom the other stereoisomer(s). Additionally, the skilled artisan knowshow to separate, enrich, and/or to selectively prepare saidstereoisomers. The compounds of the invention may be present as amixture of stereoisomers, individual stereoisomers or as an opticallyactive form.

Embodiments of the present disclosure as described in the Summary of theInvention include those described below. In the following Embodiments,reference to “a compound of Formula I” includes the definitions ofsubstituents specified in the Summary of the Invention unless furtherdefined in the Embodiments.

Embodiment 1. A compound of Formula (I):

wherein A is C₆-C₂₀ aryl optionally fused with C₁-C₂₀ heterocyclyl, orC₁-C₂₀ heteroaryl optionally fused with C₁-C₂₀ heterocyclyl, and A isoptionally substituted with one or more C₁-C₁₀ alkyl, C₆-C₂₀ aryl,halogen, OR₁₁, C(O)R₁₁, C(O)OR₁₁, NR₁₁R₁₂, SO₂R₁₁, or SO₂(OR₁₁); B isC₁-C₁₀ alkyl, C₆-C₂₀ aryl, C₁-C₂₀ heteroaryl or C₁-C₂₀ heterocyclyl andB is optionally substituted with one or more C₁-C₁₀ alkyl, C₆-C₂₀ aryl,C₁-C₂₀ heterocyclyl, halogen, OR₂₁, C(O)R₂₁, C(O)OR₂₁, NR₂₁R₂₂, SO₂R₂₁,or SO₂(OR₂₁), wherein said C₁-C₁₀ alkyl is optionally substituted withone or more halogen; R₁₁, R₁₂, R₂₁ and R₂₂ independently are H, O,C₁-C₁₀ alkyl, or C₆-C₂₀ aryl, wherein said C₁-C₁₀ alkyl is optionallysubstituted with one or more halogen;

is a single bond or double bond; n is 0 or 1; X is N or C; Y is H, OH orNH₂; Z is nil or C(O); R₃ and R₄ independently are nil, H, C₁-C₁₀ alkylor C₆-C₂₀ aryl; and R₅ is nil, H or C₁-C₁₀ alkyl.

Embodiment 2. A compound of Formula I, wherein A is C₆-C₂₀ aryloptionally fused with C₁-C₂₀ heterocyclyl, or C₁-C₂₀ heteroaryloptionally fused with C₁-C₂₀ heterocyclyl.

Embodiment 3. A compound of Formula I, wherein A is C₆-C₂₀ aryloptionally fused with C₁-C₂₀ heterocyclyl, or C₁-C₂₀ heteroaryloptionally fused with C₁-C₂₀ heterocyclyl, and A is optionallysubstituted with one or more C₁-C₁₀ alkyl, C₆-C₂₀ aryl, halogen, OR₁₁,C(O)R₁₁, C(O)OR₁₁, NR₁₁R₁₂, SO₂R₁₁, or SO₂(OR₁₁).

Embodiment 4. A compound of Formula I, wherein A is C₆-C₂₀ aryl.

Embodiment 5. A compound of Formula I, wherein A is C₆-C₂₀ aryl fusedwith C₁-C₂₀ heterocyclyl.

Embodiment 6. A compound of Formula I, wherein A is C₁-C₂₀ heteroaryl.

Embodiment 7. A compound of Formula I, wherein A is C₁-C₂₀ heteroarylfused with C₁-C₂₀ heterocyclyl.

Embodiment 8. A compound of Formula I, wherein A is C₆-C₂₀ arylsubstituted with one or more C₁-C₁₀ alkyl, C₆-C₂₀ aryl, halogen, OR₁₁,C(O)R₁₁, C(O)OR₁₁, NR₁₁R₁₂, SO₂R₁₁, or SO₂(OR₁₁).

Embodiment 9. A compound of Formula I, wherein A is C₆-C₂₀ aryl fusedwith C₁-C₂₀ heterocyclyl, and further substituted with one or moreC₁-C₁₀ alkyl, C₆-C₂₀ aryl, halogen, OR₁₁, C(O)R₁₁, C(O)OR₁₁, NR₁₁R₁₂,SO₂R₁₁, or SO₂(OR₁₁).

Embodiment 10. A compound of Formula I, wherein A is C₁-C₂₀ heteroarylsubstituted with one or more C₁-C₁₀ alkyl, C₆-C₂₀ aryl, halogen, OR₁₁,C(O)R₁₁, C(O)OR₁₁, NR₁₁R₁₂, SO₂R₁₁, or SO₂(OR₁₁).

Embodiment 11. A compound of Formula I, wherein A is C₁-C₂₀ heteroarylfused with C₁-C₂₀ heterocyclyl, and further substituted with one or moreC₁-C₁₀ alkyl, C₆-C₂₀ aryl, halogen, OR₁₁, C(O)R₁₁, C(O)OR₁₁, NR₁₁R₁₂,SO₂R₁₁, or SO₂(OR₁₁).

Embodiment 12. A compound of Formula I, wherein B is C₁-C₁₀ alkyl,C₆-C₂₀ aryl, C₁-C₂₀ heteroaryl or C₁-C₂₀ heterocyclyl.

Embodiment 13. A compound of Formula I, wherein B is C₁-C₁₀ alkyl,C₆-C₂₀ aryl or C₁-C₂₀ heteroaryl.

Embodiment 14. A compound of Formula I, wherein B is C₁-C₁₀ alkyl,C₁-C₂₀ heteroaryl or C₁-C₂₀ heterocyclyl.

Embodiment 15. A compound of Formula I, wherein B is C₁-C₁₀ alkyl,C₆-C₂₀ aryl or C₁-C₂₀ heterocyclyl.

Embodiment 16. A compound of Formula I, wherein B is C₁-C₁₀ alkyl orC₆-C₂₀ aryl.

Embodiment 17. A compound of Formula I, wherein B is C₁-C₁₀ alkyl orC₁-C₂₀ heteroaryl.

Embodiment 18. A compound of Formula I, wherein B is C₁-C₁₀ alkyl orC₁-C₂₀ heterocyclyl.

Embodiment 19. A compound of Formula I, wherein B is C₆-C₂₀ aryl orC₁-C₂₀ heteroaryl.

Embodiment 20. A compound of Formula I, wherein B is C₆-C₂₀ aryl orC₁-C₂₀ heterocyclyl.

Embodiment 21. A compound of Formula I, wherein B is C₁-C₂₀ heteroarylor C₁-C₂₀ heterocyclyl.

Embodiment 22. A compound of Formula I, wherein B is C₁-C₁₀ alkyl.

Embodiment 23. A compound of Formula I, wherein B is C₆-C₂₀ aryl.

Embodiment 24. A compound of Formula I, wherein B is C₁-C₂₀ heteroaryl.

Embodiment 25. A compound of Formula I, wherein B is C₁-C₂₀heterocyclyl.

Embodiment 26. A compound of any one of Embodiments 12-25, wherein B issubstituted with one or more C₁-C₁₀ alkyl, C₆-C₂₀ aryl, C₁-C₂₀heterocyclyl, halogen, OR₂₁, C(O)R₂₁, C(O)OR₂₁, NR₂₁R₂₂, SO₂R₂₁, orSO₂(OR₂₁).

Embodiment 27. A compound of Embodiment 26, wherein B is substitutedwith one or more C₁-C₁₀ alkyl and said C₁-C₁₀ alkyl is substituted withone or more halogen.

Embodiment 28. A compound of any one of Embodiments 8-11 and 26-27,wherein R₁₁, R₁₂, R₂₁ and R₂₂ independently are H, O, C₁-C₁₀ alkyl, orC₆-C₂₀ aryl,

Embodiment 29. A compound of Embodiment 28, wherein R₁₁, R₁₂, R₂₁ andR₂₂ independently is C₁-C₁₀ alkyl and said C₁-C₁₀ alkyl is substitutedwith one or more halogen.

Embodiment 30. A compound of Formula I, wherein n is 0.

Embodiment 31. A compound of Formula I, wherein n is 1.

Embodiment 32. A compound of Formula I, wherein X is N or C.

Embodiment 33. A compound of Formula I, wherein X is N and Y is H, OH orNH₂.

Embodiment 34. A compound of Formula I, wherein X is N and Y is H.

Embodiment 35. A compound of Formula I, wherein X is C and Y is H, OH orNH₂.

Embodiment 36. A compound of Formula I, wherein X is C and Y is H.

Embodiment 37. A compound of Formula I, wherein X is C and Y is OH.

Embodiment 38. A compound of Formula I, wherein X is C and Y is NH₂.

Embodiment 39. A compound of Formula I, wherein Z is nil or C(O)

Embodiment 40. A compound of Formula I, wherein Z is nil.

Embodiment 41. A compound of Formula I, wherein Z is C(O).

Embodiment 42. A compound of Formula I, wherein R₃ and R₄ independentlyare nil, H, C₁-C₁₀ alkyl or C₆-C₂₀ aryl.

Embodiment 43. A compound of Formula I, wherein R₃ is nil and R₄ is nil,H, C₁-C₁₀ alkyl or C₆-C₂₀ aryl.

Embodiment 44. A compound of Formula I, wherein R₃ is nil and R₄ is nil.

Embodiment 45. A compound of Formula I, wherein R₃ is nil and R₄ is H.

Embodiment 46. A compound of Formula I, wherein R₃ is nil and R₄ isC₁-C₁₀ alkyl.

Embodiment 47. A compound of Formula I, wherein R₃ is nil and R₄ isC₆-C₂₀ aryl.

Embodiment 48. A compound of Formula I, wherein R₃ is H and R₄ is nil,H, C₁-C₁₀ alkyl or C₆-C₂₀ aryl.

Embodiment 49. A compound of Formula I, wherein R₃ is H and R₄ is nil.

Embodiment 50. A compound of Formula I, wherein R₃ is H and R₄ is H.

Embodiment 51. A compound of Formula I, wherein R₃ is H and R₄ is C₁-C₁₀alkyl.

Embodiment 52. A compound of Formula I, wherein R₃ is H and R₄ is C₆-C₂₀aryl.

Embodiment 53. A compound of Formula I, wherein R₃ is C₁-C₁₀ alkyl andR₄ is nil, H, C₁-C₁₀ alkyl or C₆-C₂₀ aryl.

Embodiment 54. A compound of Formula I, wherein R₃ is C₁-C₁₀ alkyl andR₄ is nil.

Embodiment 55. A compound of Formula I, wherein R₃ is C₁-C₁₀ alkyl andR₄ is H.

Embodiment 56. A compound of Formula I, wherein R₃ is C₁-C₁₀ alkyl andR₄ is C₁-C₁₀ alkyl.

Embodiment 57. A compound of Formula I, wherein R₃ is C₁-C₁₀ alkyl andR₄ is C₆-C₂₀ aryl.

Embodiment 58. A compound of Formula I, wherein R₃ is C₆-C₂₀ aryl and R₄is nil, H, C₁-C₁₀ alkyl or C₆-C₂₀ aryl.

Embodiment 59. A compound of Formula I, wherein R₃ is C₆-C₂₀ aryl and R₄is nil.

Embodiment 60. A compound of Formula I, wherein R₃ is C₆-C₂₀ aryl and R₄is H.

Embodiment 61. A compound of Formula I, wherein R₃ is C₆-C₂₀ aryl and R₄is C₁-C₁₀ alkyl.

Embodiment 62. A compound of Formula I, wherein R₃ is C₆-C₂₀ aryl and R₄is C₆-C₂₀ aryl.

Embodiment 63. A compound of Formula I, wherein R₅ is nil, H or C₁-C₁₀alkyl.

Embodiment 64. A compound of Formula I, wherein R₅ is nil or H.

Embodiment 65. A compound of Formula I, wherein R₅ is nil, or C₁-C₁₀alkyl.

Embodiment 66. A compound of Formula I, wherein R₅ is H or C₁-C₁₀ alkyl.

Embodiment 67. A compound of Formula I, wherein R₅ is nil.

Embodiment 68. A compound of Formula I, wherein R₅ is H.

Embodiment 69. A compound of Formula I, wherein R₅ is C₁-C₁₀ alkyl.

Embodiments of this invention, including Embodiments 1-69 above as wellas any other embodiments described herein, can be combined in anymanner, and the descriptions of variables in the embodiments pertain notonly to the compounds of Formula I but also to the starting compoundsand intermediate compounds useful for preparing the compounds of FormulaI. In addition, embodiments of this invention, including Embodiments1-69 above as well as any other embodiments described herein, and anycombination thereof, pertain to the compositions and methods of thepresent disclosure.

Combinations of Embodiments 1-69 are illustrated by:

Embodiment A. A compound of Formula (I)

wherein

-   -   A is C₆-C₂₀ aryl optionally fused with C₁-C₂₀ heterocyclyl, or        C₁-C₂₀ heteroaryl optionally fused with C₁-C₂₀ heterocyclyl, and        A is optionally substituted with one or more C₁-C₁₀ alkyl,        C₆-C₂₀ aryl, halogen, OR₁₁, C(O)R₁₁, C(O)O_(Rn), NR₁₁R₁₂,        SO₂R₁₁, or SO₂(OR₁₁);    -   B is C₁-C₁₀ alkyl, C₆-C₂₀ aryl, C₁-C₂₀ heteroaryl or C₁-C₂₀        heterocyclyl, and B is optionally substituted with one or more        C₁-C₁₀ alkyl, C₆-C₂₀ aryl, C₁-C₂₀ heterocyclyl, halogen, OR₂₁,        C(O)R₂₁, C(O)OR₂₁, NR₂₁R₂₂, SO₂R₂₁, or SO₂(OR₂₁), wherein said        C₁-C₁₀ alkyl is optionally substituted with one or more halogen;    -   R₁₁, R₁₂, R₂₁ and R₂₂ independently are H, O, C₁-C₁₀ alkyl, or        C₆-C₂₀ aryl, wherein said C₁-C₁₀ alkyl is optionally substituted        with one or more halogen;    -   is a single bond or double bond;    -   n is 0 or 1;    -   X is N or C;    -   Y is H, OH or NH₂;    -   Z is nil or C(O);    -   R₃ and R₄ independently are nil, H, C₁-C₁₀ alkyl or C₆-C₂₀ aryl;        and    -   R₅ is nil, H or C₁-C₁₀ alkyl.

Embodiment B. A compound of Embodiment A represented by Formula (II)

wherein

-   -   A is C₆-C₂₀ aryl optionally fused with C₁-C₂₀ heterocyclyl, or        C₁-C₂₀ heteroaryl optionally fused with C₁-C₂₀ heterocyclyl, and        A is optionally substituted with one or more C₁-C₁₀ alkyl,        C₆-C₂₀ aryl, halogen, OR₁₁, C(O)R₁₁, C(O)O_(Rn), NR₁₁R₁₂,        SO₂R₁₁, or SO₂(OR₁₁);    -   B is C₁-C₁₀ alkyl, C₆-C₂₀ aryl, C₁-C₂₀ heteroaryl or C₁-C₂₀        heterocyclyl and is optionally substituted with one or more        C₁-C₁₀ alkyl, C₆-C₂₀ aryl, C₁-C₂₀ heterocyclyl, halogen, OR₂₁,        C(O)R₂₁, C(O)OR₂₁, NR₂₁R₂₂, SO₂R₂₁, or SO₂(OR₂₁), wherein said        C₁-C₁₀ alkyl is optionally substituted with one or more halogen;    -   R₁₁, R₁₂, R₂₁ and R₂₂ independently are H, O, C₁-C₁₀ alkyl, or        C₆-C₂₀ aryl, wherein said C₁-C₁₀ alkyl is optionally substituted        with one or more halogen;    -   Y is OH or NH₂;    -   Z is nil or C(O); and    -   R₅ is nil, H, or C₁-C₁₀ alkyl.

Embodiment C. A compound of Embodiment A represented by Formula (III)

wherein

-   -   A is C₆-C₂₀ aryl optionally fused with C₁-C₂₀ heterocyclyl, or        C₁-C₂₀ heteroaryl optionally fused with C₁-C₂₀ heterocyclyl, and        A is optionally substituted with one or more C₁-C₁₀ alkyl,        C₆-C₂₀ aryl, halogen, OR₁₁, C(O)R₁₁, C(O)O_(Rn), NR₁₁R₁₂,        SO₂R₁₁, or SO₂(OR₁₁);    -   B is C₁-C₁₀ alkyl, C₆-C₂₀ aryl, C₁-C₂₀ heteroaryl or C₁-C₂₀        heterocyclyl, and B is optionally substituted with one or more        C₁-C₁₀ alkyl, C₆-C₂₀ aryl, C₁-C₂₀ heterocyclyl, halogen, OR₂₁,        C(O)R₂₁, C(O)OR₂₁, NR₂₁R₂₂, SO₂R₂₁, or SO₂(OR₂₁), wherein said        C₁-C₁₀ alkyl is optionally substituted with one or more halogen;    -   R₁₁, R₁₂, R₂₁ and R₂₂ independently are H, O, C₁-C₁₀ alkyl, or        C₆-C₂₀ aryl, wherein said C₁-C₁₀ alkyl is optionally substituted        with one or more halogen;    -   Z is nil or C(O); and    -   each R₃ and R₄ independently is nil, H, C₁-C₁₀ alkyl or C₆-C₂₀        aryl.

Embodiment D. A compound of any one of Embodiments A-C wherein

-   -   A is substituted or unsubstituted C₆-C₂₀ aryl or C₆-C₂₀ aryl        fused with C₁-C₂₀ heterocyclyl.

Embodiment E. A compound of any one of Embodiments A-D wherein

-   -   A is phenyl group substituted with one or more methoxy group,        (dimethylamide) phenyl group, Br, OR₁₁, or C(O)OR₁₁, wherein R₁₁        is H, O, C₁-C₁₀ alkyl, or C₆-C₂₀ aryl, wherein said C₁-C₁₀ alkyl        is optionally substituted with one or more halogen.

Embodiment F. A compound of any one of Embodiments A-C wherein

-   -   A is substituted or unsubstituted C₁-C₂₀ heteroaryl having at        least one heteroatom selected from S, N, and 0.

Embodiment G. A compound of Embodiment F. wherein

-   -   A is pyrroline, furan, thiophene, pyridine, pyran, or thiopyran.

Embodiment H. A compound of any one of Embodiments A-D wherein

-   -   A is substituted or unsubstituted C₆-C₂₀ aryl fused with C₁-C₂₀        heterocyclyl having at least one heteroatom selected from S, N,        and O.

Embodiment I. A compound of Embodiment H wherein

-   -   A is 1,3-benzodioxole, indoline, indole, indazole, benzofuran,        benzo[c]thiophene, benzo[b]thiophene, 1,2-benzisoxazole,        1,2-benzisothiazole, 2,1-benzisothiazole, benzoxazole, or        benzthiazole.

Embodiment J. A compound of any one of Embodiments A-I wherein

-   -   B is substituted or unsubstituted C₁-C₁₀ alkyl.

Embodiment K. A compound of Embodiment J wherein

-   -   B is methyl, ethyl, n-propyl, i-propyl or tert-butyl group.

Embodiment L. A compound of any one of Embodiments A-I wherein

-   -   B is substituted or unsubstituted C₆-C₂₀ aryl.

Embodiment M. A compound of Embodiment L wherein

-   -   B is a phenyl group substituted with one or more methyl, CF₃,        halogen, pyrrolidine, OR₂₁, C(O)R₂₁, NR₂₁R₂₂, or SO₂R₂₁.

Embodiment N. A compound of Embodiment M wherein

-   -   R₂₁ is H, O, methyl, ethyl, n-propyl, or CF₃; and R₂₂ is H or 0.

Embodiment O. A compound of any one of Embodiments A-I wherein

-   -   B is substituted or unsubstituted C₁-C₂₀ heteroaryl.

Embodiment P. A compound of Embodiment 0 wherein

-   -   B is pyrroline, furan, thiophene, pyridine, pyran, or thiopyran.

Embodiment Q. A compound of any one of Embodiments A-P wherein

-   -   n is 0.

Embodiment R. A compound of any one of Embodiments A-Q wherein

-   -   X is C.

Embodiment S. A compound of any one of Embodiments A-R wherein

-   -   Y is H, OH or NH₂.

Embodiment T. A compound of any one of Embodiments A-Q wherein

-   -   X is N.

Embodiment U. A compound of Embodiment T wherein

-   -   Y is H or NH₂.

Embodiment V. A compound of any one of Embodiments A-U wherein

-   -   Z is nil or C(O).

Embodiment W. A compound of Embodiments A-P wherein

-   -   n is 1.

Embodiment X. A compound of Embodiment W wherein

-   -   R₃ and R₄ independently are nil, H, methyl, ethyl, n-propyl,        i-propyl, tert-butyl, phenyl, benzyl, or naphthyl.

Embodiment Y. A compound of Embodiment A wherein

-   -   R₅ is nil, H, methyl, ethyl, n-propyl, i-propyl or tert-butyl        group.

Specific embodiments include compounds of Formula I selected from agroup consisting of:

Also noteworthy as embodiments of the present disclosure arecompositions comprising a compound of any of the preceding Embodiments,as well as any other embodiments described herein, and any combinationsthereof. The composition can be formulated to any form of formulationthat is considered to be safe, effective, and convenient for use.Preferably, the pharmaceutical composition may further comprise apharmaceutically acceptable component as defined in the previousparagraphs. A usable pharmaceutically acceptable salt, carrier, orexcipient are disclosed in various references including Handbook ofPharmaceuticals Excipients edited by Raymond C Rowe, Paul J Sheskey, andMarian E Quinn. In a unlimited embodiment, said pharmaceuticallyacceptable carrier or excipient can be selected from the groupconsisting of inert diluents, dispersing and/or granulating agents,surface active agents and/or emulsifiers, disintegrating agents, bindingagents, preservatives, buffering agents, lubricating agents and oils.Said compositions optionally further comprising at least one additionalbiologically active compound or agent.

In a particular embodiment, said composition could be a pharmaceuticalcomposition. The pharmaceutical composition might comprise but notlimited to a single unit dose of the active ingredient (for instance,the compound of the present disclosure). For purposes of treatment, adose unit can be in the form of a discrete article such as but notlimited to a tablet or capsule, or can be a measurable volume of asolution, suspension or the like containing a unit dose of the activeingredient. The term “unit dose” herein refers to an amount of activeingredient intended for a single but not limited to oral, intravenous,intramuscular, cutaneous, subcutaneous, intrathecal, transdermal,implantation, sublingual, buccal, rectal, vaginal, ocular, otic, nasal,inhalation, or nebulization administration to a subject for inhibitingNADPH oxidase (NOX). The inhibitory to NOX may require periodicadministration of unit doses of the compound of the present disclosure,for example one unit dose two or more times a day, one unit dose witheach meal, one unit dose every four hours or other interval, or only oneunit dose per day.

Further noteworthy as embodiments of the present disclosure arecompositions for inhibiting NADPH oxidase (NOX) activity comprising acompound of any of the preceding Embodiments, as well as any otherembodiments described herein, and any combinations thereof, and at leastone of a pharmaceutically acceptable excipient or carrier selected fromthe group consisting of inert diluents, dispersing and/or granulatingagents, surface active agents and/or emulsifiers, disintegrating agents,binding agents, preservatives, buffering agents, lubricating agents,and/or oils, said compositions optionally further comprising at leastone additional biologically active compound or agent. Embodiments of theinvention further include methods for inhibiting NADPH oxidase (NOX)activity in cell comprising administering to the cell an effectiveamount of the compound or the pharmaceutical composition of any of thepreceding Embodiments (e.g., as a composition described herein).

Examples

One or more of the following methods and variations as described inSchemes I-XII can be used to prepare the compounds of Formula I. Thedefinitions of substituents in the compounds below are as defined aboveunless otherwise noted. The following abbreviations are used: DCM isdichloromethane, DMF is N,N-dimethylformamide, DMSO is dimethylsulfoxide, EA is ethyl acetate, MeOH is methanol, TFA is trifluoroaceticacid, and THF is tetrahydrofuran. ¹H NMR spectra are reported in ppmdownfield from tetramethylsilane; “s” means singlet, “d” means doublet,“t” means triplet, “q” means quartet, “m” means multiplet, “dd” meansdoublet of doublets, “dt” means doublet of triplets, and “br s” meansbroad singlet.

A. Synthesis of Compounds

Variations on the following general synthetic methods will be readilyapparent to those of ordinary skill in the art and are deemed to bewithin the scope of the present disclosure.

A-1. Synthesis of Compounds 2-20 and 24-27

As explained in details below, Compounds 2-20 and 24-27 are synthesizedby the method depicted in Scheme I below.

Step 1: Preparation of 2-(4-methoxyphenyl)-2-oxoethyl2-amino-4,5-dimethoxybenzoate

2-amino-4,5-dimethoxylbenzoic acid (1.97 g, 10 mmol) was dissolved inDMF (20 mL), and to the solution was added potassium carbonate (1.38, 10mmol). The reaction mixture was heated at 90° C. and stirred for 1 hr.Then the solution was cooled to 20° C. and p-methoxy-2-bromoacetophenone(2.4 g, 10.5 mmol) was added. The slightly exothermic reaction tookplace and the temperature increased to 25° C. to 32° C. in 5 minutes.After stirring for 30 minutes the reaction mixture was heated to 50° C.and kept at this temperature for 30 minutes. The content of the flaskwas then poured into a 100 g mixture of water and ice. The precipitatedsolid material was collected by filtration, washed with water and dried.The dried solid was further recrystallized with ethanol to give Compound1 (2-(4-methoxyphenyl)-2-oxoethyl 2-amino-4,5-dimethoxybenzoate) with ayield of 3.3 g (95%).

Step 2: Preparation of 3-hydroxy-6,7-dimethoxy-2-(4-methoxyphenyl)quinolin-4(1H)-one 2,2,2-trifluoroacetate

A solution of Compound 1 (0.6 g, 1.74 mmol) in TFA was heated to refluxfor overnight, until reaction completed. Furthermore, the reactionmixture was cooled to room temperature, and TFA was removed in vacuo.The residue was added ice-water and stirred for 30 minutes. The solidwas collected and washed by H₂O and methanol to obtain Compound 2(3-hydroxy-6,7-dimethoxy-2-(4-methoxyphenyl) quinolin-4(1H)-one2,2,2-trifluoroacetate) with a yield of 0.5 g (87%).

Step 3: Preparation of 3,6,7-trihydroxy-2-(4-hydroxyphenyl)quinolin-4(1H)-one hydrobromide

A suspension of Compound 2 (0.2 g, 0.6 mmol) in 20 mL HBr (47%) washeated to 100° C. under an atmosphere of dry nitrogen for 23 hr. Thesolvents were evaporated under reduced pressure and the product wascrystallized (by MeOH/EA=1:3). Compound 3(3,6,7-trihydroxy-2-(4-hydroxyphenyl) quinolin-4(1H)-one hydrobromide)is obtained with a yield of 25%.

Similarly, by replacing the p-methoxy-2-bromoacetophenone in Step 1,Compounds 4-20 and 24-27 can be prepared by the steps described above.

A-2. Synthesis of Compounds 21 and 22 Step 1A: Preparation of2-bromo-1-(2-ethoxyphenyl) ethan-1-one

To a suspension of 1-(2-hydroxyphenyl)ethan-1-one (2.0 g, 14.69 mmol),anhydrous K₂CO₃ (2.6 g, 18.8 mmol) and in DMF (80 ml) was addedbromoethane (5 mL, 67 mmol). The mixture was heated at 50° C. until thestarting ketone was completely consumed. After cooling to roomtemperature, the mixture was filtered, and the solvent was removed byrotary evaporation. The residue was purified by flash chromatography.The product is 1-(2-ethoxyphenyl) ethan-1-one with n-hexane/EA (9:1) aseluent, in the presence of colorless oil. The yield is 95%.

In addition, 1-(2-ethoxyphenyl) ethan-1-one (2.55 g, 15.53 mmol) wasdissolved in Et₂O (40 mL) and bromine (0.7 mL, 14 mmol) was addeddropwise while stirring stir the mixture in the dark at ambienttemperature. Wash the reaction mixture with a saturated aqueous Na₂CO₃solution. The product is dried over MgSO₄, filtered, collected thefiltrate, and removed the volatiles under reduced pressure to providethe compound 2-bromo-1-(2-ethoxyphenyl) ethan-1-one, which was purifiedby flash chromatography (n-hexane/EA=9:1) with a yield of 1.8 g (48%).

The obtained compound were then used in Steps 1-3 of A-1 in replace ofthe p-methoxy-2-bromoacetophenone to prepare Compounds 21 and 22.

A-3. Synthesis of Compound 23 Step 1A: Preparation of2-bromo-1-(2-(methylsulfonyl)phenyl) ethan-1-one

A mixture of 1-(2-iodophenyl) ethan-1-one (1.0 g, 4.06 mmol), sodiummethanesulfunate (0.83 g, 8.12 mmol) and copper iodide (77 mg, 0.4 mmol)in 10 mL of DMSO was heated to 100° C. under argon. The cooled mixturewas partitioned by EA and water. The organic layer was separated, andthe aqueous layer was extracted with EA twice. The combined organiclayers were washed with brine, dried over MgSO₄, and concentrated invacuo to afford the product, 1-(2-(methylsulfonyl)phenyl) ethan-1-one,which is yellow solid with a yield of 0.77 g (95%).

To the stirred mixture copper (II) bromide (0.22 g, 0.98 mmol) in EA (6ml) was added solution of 1-(2-(methylsulfonyl)phenyl)ethan-1-one (0.1g, 0.5 mmol) in EA (10 ml) dropwise under the room temperature. Afteraddition was finished, the mixture was heated at 80° C. for 18 hours.After that, the mixture was cooled to room temperature, filtered andprecipitate was washed by EA (10 ml). The EA solution was treated withwater, the organic phase was separated and dried over Na₂SO₄, andconcentrated in vacuo. The crude product,2-bromo-1-(2-(methylsulfonyl)phenyl) ethan-1-one, which was purified bycolumn chromatography (DCM/Hexanes=2/1) to give the purified productwith a yield of 0.115 g (82%).

The obtained compound were then used in Steps 1-3 of A-1 in replace ofthe p-methoxy-2-bromoacetophenone to prepare Compound 23.

A-4. Synthesis of Compounds 29 and 30

Preparation of2-(2-aminophenyl)-3-hydroxy-6,7-dimethoxyquinolin-4(1H)-one

By following the same methods as in Scheme I, Compound 29(3-hydroxy-6,7-dimethoxy-2-(2-nitrophenyl) quinolin-4(1H)-one2,2,2-trifluoroacetate) was obtained. To a solution of Compound 29 (0.3g, 0.66 mmol) in ethanol (12 mL) was added hydrazine hydrate (50%) (0.4mL, 6.6 mmol) and an excess of Raney nickel slurry in water (1 mmol) wasadded slowly. The reaction was stirred at 70° C. for 1 hr. After 1 hr,the bubbling ceased, and the mixture was cooled to room temperature andfiltered through Celite. The filtrate was condensed under reducedpressured and the residue was washed with H₂O/MeOH to afford the desiredproduct 30 (2-(2-aminophenyl)-3-hydroxy-6,7-dimethoxyquinolin-4(1H)-one)(0.19 g, 93%).

A-5. Synthesis of Compounds 32 and 33

Step 1: Preparation of bis(2-(4-methoxyphenyl)-2-oxoethyl)2-aminoterephthalate

To the solution of 2-Aminoterephthalic acid (0.91 g, 5 mmol) in DMF (20mL) was added potassium carbonate (1.38 g, 10 mmol) andp-methoxy-2-bromoacetophenone (2.4 g, 10.5 mmol). The reaction mixturewas heated at 50° C. and stirred for 18 hr. Then the content of theflask was then poured into a 100 g mixture of water and ice. Theprecipitated solid material was collected by filtration, washed withwater and methanol to afford Compound 31(bis(2-(4-methoxyphenyl)-2-oxoethyl) 2-aminoterephthalate) with a yieldof 1.9 g (80%).

Step 2: Preparation of 2-(4-methoxyphenyl)-2-oxoethyl3-hydroxy-2-(4-methoxyphenyl)-4-oxo-1,4-dihydroquinoline-7-carboxylate2,2,2-trifluoroacetate

A solution of Compound 31 (0.5 g, 1.1 mmol) in TFA (12 mL) was heated toreflux for overnight, until reaction completed. Then the reactionmixture was cooled to room temperature, and TFA was removed in vacuo.The residue was added ice-water and stirred for 30 minutes. The solidwas collected and washed by H₂O and methanol to obtain Compound 32(2-(4-methoxyphenyl)-2-oxoethyl3-hydroxy-2-(4-methoxyphenyl)-4-oxo-1,4-dihydroquinoline-7-carboxylate2,2,2-trifluoroacetate) with a yield of 0.5 g (79%).

Step 3: Preparation of3-hydroxy-2-(4-methoxyphenyl)-4-oxo-1,4-dihydroquinoline-7-carboxylicacid

A solution of Compound 32 (0.1 g, 0.17 mmol) in THF (5 mL) was add 5 mLof 1N LiOH(aq) and stirred at room temperature for overnight, untilreaction completed. The reaction mixture was then cooled to 0° C., and2N HCl was added slowly until pH=1-2. The solid was collected and washedby H₂O and methanol to obtain Compound 33(3-hydroxy-2-(4-methoxyphenyl)-4-oxo-1,4-dihydroquinoline-7-carboxylicacid) with a yield of 23 mg.

A-6. Synthesis of Compounds 34

A solution of sodium ethoxide (Na, 2 mmol in dry EtOH, 5 mL) was add asuspension of 2,3-dihydro-2,3-dioxo-1H-indole (0.17 g, 1.0 mmol) and2-bromo-2 methoxy acetophenone (0.23 g, 1.0 mmol) in dry EtOH (5 mL).The mixture was maintained at 0-5° C. for 2 hr. The reaction mixture wasallowed to warm up to room temperature and stirred for 18 h. Then themixture was acidified with 2N HCl_((aq.)) and extracted with EA. Theorganic phase was dried with MgSO₄ and concentrated under reducedpressure to give the crude mixture. The crude mixture was purified bycolumn chromatography (EA/Hexanes=1/2) to give Compound 34(3-hydroxy-6-methoxy-2-(2-methoxybenzoyl) quinolin-4(1H)-one) with ayield of 0.1 g (30%).

A-7. Synthesis of Compounds 35-38 Step 1: Preparation of2-amino-1-(2-chlorophenyl) ethan-1-one hydrochloride

To the full dissolution of 2-bromo-1-(2-chlorophenyl) ethan-1-one (3.0g, 12.85 mmol) in the DMSO was added sodium azide (1.2 g) and stirred atroom temperature for 15 hr. The resulting mixture was diluted with H₂O.The layers are separated and extracted with EA. The combined organiclayers are dried over MgSO₄, filtered and concentrated on a rotaryevaporator to give the compound 2-azido-1-(2-chlorophenyl) ethan-1-onewith a yield of 2.3 g (11.8 mmol).

A mixture of the 2-azido-1-(2-chlorophenyl)ethan-1-one (2.3 g, 11.8mmol) and 5% Pd/C (0.5 g) in MeOH (100 mL) was added the concentratedHCl (2 mL) stirred under an atmosphere of H₂ (1 atm) at room temperaturefor 6 h. The mixture was filtered through celite and the filtrate wasconcentrated. The residue was not purified to provide the product.Compound 2-amino-1-(2-chlorophenyl) ethan-1-one hydrochloride with ayield of 2.2 g (10.73 mmol).

Step 2: Preparation of2-(2-chlorophenyl)-7,8-dimethoxy-3,4-dihydro-5H-benzo[e][1,4]diazepin-5-one and3-amino-2-(2-chlorophenyl)-6,7-dimethoxyquinolin-4(1H)-one

A mixture of the 2-amino-1-(2-chlorophenyl)ethan-1-one hydrochloride(1.0 g, 4.9 mmol), isatoic anhydride (0.9 g, 4.45 mmol), and Na₂CO₃ (0.6g, 5.34 mmol) in H₂O (20 mL) was stirred at 80° C. for 4 hr. After thetemperature cooled down to room temperature, the precipitate wascollected by filtration and washed with H₂O and ether to give theCompound 35(2-(2-chlorophenyl)-7,8-dimethoxy-3,4-dihydro-5H-benzo[e][1,4]diazepin-5-one) with a yield of 870.0 mg (2.64 mmol).

A mixture of the Compound 35(2-(2-chlorophenyl)-7,8-dimethoxy-3,4-dihydro-5H-benzo[e][1,4]diazepin-5-one) (100.0 mg, 0.30 mmol) and PPA (5 mL) was stirred at 150°C. for 2 h. After the temperature cooled down to 0° C., the resultingmixture was poured onto Na₂CO_(3(aq)), and after foaming had stopped,water was added. After 30 min of stirring, a solid product was filteredoff and washed with water and methanol to give the Compound 36(3-amino-2-(2-chlorophenyl)-6,7-dimethoxyquinolin-4(1H)-one) with ayield of 10.0 mg (0.03 mmol).

Similarly, Compounds 37-38 are synthesized by using the same methods asin Scheme VII and VIII.

A-8. Synthesis of Compound 39

Compound 39 was synthesized by Scheme IX, which are using the methodssimilar to Scheme I.

A-9. Synthesis of Compound 40

Compound 25 (0.3 g, 1.3 mmol) and 3-(N,N-dimethylamino)phenyl boronicacid (0.258 g, 1.56 mmol) were added into 3 ml ethanol, followed byadding sodium carbonate (1.1 g, 10.4 mmol) andtetrakis(triphenylphosphine)palladium(0) (0.075 g, 0.065 mmol). Thereaction mixture was heated under reflux overnight. After cooling toroom temperature, the residue was filtered. The filtrate was extractedwith EA and dried in vacuo. The dried residue was then applied withcolumn chromatography with EA: hexane (1:1) as eluent to obtain Compound40 with a yield of 0.42 g (80%).

A-10. Synthesis of Compound 41-44 Step 1: Preparation of(3-hydroxy-6,7-dimethoxy-2-(2-(trifluoromethyl) phenyl)quinolin-4(1H)-one

To the full dissolution of Compound 17 (30.0 mg, 0.06 mmol) in H₂O wasadded 1N NaOH (0.1 mL) stirred at room temperature for 3 hr. Theprecipitate was collected by filtration and washed with H₂O to give thefree base Compound 41 (3-hydroxy-6,7-dimethoxy-2-(2-(trifluoromethyl)phenyl) quinolin-4(1H)-one) with a yield of 13.0 mg (0.04 mmol).

Step 2A: Preparation of3-hydroxy-6,7-dimethoxy-2-(2-(trifluoromethyl)phenyl) quinolin-4(1H)-onehydrochloride

To the full dissolution of Compound 41 (0.29 g, 0.79 mmol) in theco-solvent of MeOH:DCM=2:1 was added 2N HCl in ether and stirred at roomtemperature for 3 hr. The precipitate was collected by filtration andwashed with ether to give the Compound 42(3-hydroxy-6,7-dimethoxy-2-(2-(trifluoromethyl)phenyl)quinolin-4(1H)-one hydrochloride) with a yield of 0.17 g (0.42 mmol).

Step 2B: Preparation of3-hydroxy-6,7-dimethoxy-2-(2-(trifluoromethyl)phenyl)quinolin-4(1H)-oneacetate

To the full dissolution of Compound 41(3-hydroxy-6,7-dimethoxy-2-(2-(trifluoromethyl) phenyl)quinolin-4(1H)-one) (28.0 mg, 0.08 mmol) in the co-solvent ofMeOH:DCM=2:1 was added acetic acid (0.01 mL) and stirred at 100° C. for15 hr. The precipitate was collected by filtration and washed with etherto give the Compound 43(3-hydroxy-6,7-dimethoxy-2-(2-(trifluoromethyl)phenyl)quinolin-4(1H)-oneacetate) with a yield of 6.0 mg (0.014 mmol).

Step 2C: Preparation of3-hydroxy-6,7-dimethoxy-2-(2-(trifluoromethyl)phenyl)quinolin-4(1H)-onemethanesulfonate

To the full dissolution of Compound 41(3-hydroxy-6,7-dimethoxy-2-(2-(trifluoromethyl)phenyl)quinolin-4(1H)-one) (75.0 mg, 0.21 mmol) in the co-solvent ofMeOH:DCM=2:1 was added methanesulfonic acid (16.3 μl) and stirred atroom temperature for 15 hr. The precipitate was collected by filtrationand washed with ether to give the Compound 44(3-hydroxy-6,7-dimethoxy-2-(2-(trifluoromethyl)phenyl)quinolin-4(1H)-onemethanesulfonate) with a yield of 21.0 mg (0.046 mmol).

A-11. Synthesis of Compounds 45-50

Compounds 45-50 were synthesized by using the methods as in Scheme XI.

A-12. Synthesis of Compound 51-52

Compounds 51-52 were synthesized by using the methods as in Scheme XII.

B. Pharmaceutically Acceptable Salts

The term “active agent” as used herein, includes the pharmaceuticallyacceptable salts of the compound. Pharmaceutically acceptable salts aresalts that retain the desired biological activity of the parent compoundand do not impart undesired toxicological effects. Examples of suchsalts are (a) acid addition salts formed with inorganic acids, forexample hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoricacid, nitric acid and the like; and salts formed with organic acids suchas, for example, acetic acid, oxalic acid, tartaric acid, succinic acid,maleic acid, fumaric acid, gluconic acid, citric acid, malic acid,ascorbic acid, benzoic acid, tannic acid, palmitic acid, alginic acid,polyglutamic acid, naphthalenesulfonic acid, methanesulfonic acid,p-toluenesulfonic acid, naphthalenedisulfonic acid, polygalacturonicacid, and the like; and (b) salts formed from elemental anions such aschlorine, bromine, and iodine. In other particular embodiments,pharmaceutically acceptable salts are formed with malic acid. Inparticular embodiments, pharmaceutically acceptable salts are formedwith hydrochloric acid. Active agents used to prepare compositions forthe present disclosure may alternatively be in the form of apharmaceutically acceptable free base of active agent. Because the freebase of the compound is less soluble than the salt, free basecompositions are employed to provide more sustained release of activeagent to the target area. Active agent present in the target area whichhas not gone into solution is not available to induce a physiologicalresponse, but serves as a depot of bioavailable drug which graduallygoes into solution.

INDEX TABLE OF THE COMPOUNDS Cmpd. No. and Structure ¹H NMR Data

¹H-NMR (300 MHz, DMSO): δ 11.25 (s, 1H), 7.78-7.74 (d, 2H), 7.41 (s,1H), 7.18 (s, 1H), 7.13-7.09 (d, 2H), 3.85 (s, 6H), 3.84 (s, 3H). ESI-MSm/z calcd for C₂₀H₁₈F₃NO₇, 441.10 found 328.1 [M + H]⁺ Chemical Formula:C₂₀H₁₈F₃NO₇ Exact Mass: 441.10 Molecular Weight: 441.36 Compound 2

¹H-NMR (400 MHz, DMSO): δ 7.72-7.70 (d, 2H), 7.51 (s, 1H), 7.30 (s, 1H),7.03-7.01 (d, 2H). ESI-MS m/z calcd for C₁₅H₁₂BrNO₅ 364.99, found 286.1[M + H]⁺ Chemical Formula: C₁₅H₁₂BrNO₅ Exact Mass: 364.99 MolecularWeight: 366.17 Compound 3

¹H-NMR (300 MHz, DMSO): δ 7.52-7.50 (d, 1H), 7.47 (s, 1H), 7.40 (s, 1H),7.38-7.36 (m, 1H), 7.25 (s, 1H), 7.13-7.10 (dd, 1H), 3.89- 3.84 (m, 6H),3.56 (s, 3H). ESI-MS m/z calcd for C₂₀H₁₈F₃NO₇ 441.10, found 328.3 [M +H]⁺ Chemical Formula: C₂₀H₁₈F₃NO₇ Exact Mass: 441.10 Molecular Weight:441.36 Compound 4

¹H-NMR (400 MHz, DMSO): δ 7.54-7.50 (m, 1H), 7.46 (s, 1H), 7.43-7.41 (d,1H), 7.22-7.20 (d, 1H), 7.12-7.08 (m, 1H), 7.07 (s, 1H), 3.88 (s, 3H),3.85 (s, 3H), 3.78 (s, 3H). ESI-MS m/z calcd for C₂₀H₁₈F₃NO₇ 441.10,found 328.3 [M + H]⁺ Chemical Formula: C₂₀H₁₈F₃NO₇ Exact Mass: 441.10Molecular Weight: 441.36 Compound 5

¹H-NMR (400 MHz, DMSO): δ 11.27 (br, 1H), 7.71 (d, 2H), 7.42 (s, 1H),7.36 (d, 2H), 7.19 (s, 1H), 3.86 (s, 6H), 2.40 (s, 3H). ESI-MS m/z calcdfor C₂₀H₁₈F₃NO₆ 425.11, found 312.3 [M + H]⁺ Chemical Formula:C₂₀H₁₈F₃NO₆ Exact Mass: 425.11 Molecular Weight: 425.36 Compound 6

¹H-NMR (400 MHz, DMSO): δ 8.00 (s, 1H), 7.87-7.85 (m, 1H), 7.45 (s, 1H),7.33- 7.29 (m, 2H), 3.90 (s, 3H), 3.84 (s, 3H). ESI-MS m/z calcd forC₁₇H₁₄F₃NO₆S 417.05, found 304.2 [M + H]⁺ Chemical Formula: C₁₇H₁₄F₃NO₆SExact Mass: 417.05 Molecular Weight: 417.36 Compound 7

¹H-NMR (400 MHz, DMSO): δ 7.85-7.82 (d, 2H), 7.65-7.63 (d, 2H), 7.46 (s,1H), 7.20 (s, 1H), 3.87 (s, 6H). ESI-MS m/z calcd for C₁₉H₁₅ClF₃NO₆445.05, found 332.2 [M + H]⁺ Chemical Formula: C₁₉H₁₅ClF₃NO₆ Exact Mass:445.05 Molecular Weight: 445.78 Compound 8

¹H-NMR (400 MHz, DMSO): δ 7.66-7.48 (m, 4H), 7.45 (s, 1H), 6.98 (s, 1H),3.86 (s, 3H), 3.83 (s, 3H). ESI-MS m/z calcd for C₁₉H₁₅ClF₃NO₆ 445.05,found 332.3 [M + H]⁺. Chemical Formula: C₁₉H₁₅ClF₃NO₆ Exact Mass: 445.05Molecular Weight: 445.78 Compound 9

¹H-NMR (300 MHz, DMSO): δ 7.92-7.90 (d, 1H), 7.73-7.70 (d, 2H),7.24-7.22 (d, 1H), 7.12- 7.09 (d, 2H), 3.83 (s, 3H). ESI-MS m/z calcdfor C₁₆H₁₂F₃NO₅S 387.04, found 274.2 [M + H]⁺ Chemical Formula:C₁₆H₁₂F₃NO₅S Exact Mass: 387.04 Molecular Weight: 387.33 Compound 10

¹H-NMR (400 MHz, DMSO): δ 13.26 (br, 1H), 7.78 (d, 2H), 7.54 (s, 1H),7.49 (s, 1H), 7.73 (d, 2H), 3.93 (s, 6H), 3.36-3.31 (m, 4H), 2.02- 1.91(m, 4H). ESI-MS m/z calcd for C₂₃H₂₃F₃N₂O₆ 480.15, found 367.4 [M + H]⁺Chemical Formula: C₂₃H₂₃F₃N₂O₆ Exact Mass: 480.15 Molecular Weight:480.44 Compound 11

¹H-NMR (300 MHz, DMSO): δ 8.03 (d, 2H), 7.93 (d, 2H), 7.43 (s, 1H), 7.15(s, 1H), 3.87 (s, 6H). ESI-MS m/z calcd for C₂₀H₁₅F₆NO₆ 479.08, found366.3 [M + H]⁺ Chemical Formula: C₂₀H₁₅F₆NO₆ Exact Mass: 479.08Molecular Weight: 479.33 Compound 12

¹H-NMR (300 MHz, DMSO): δ 7.88-7.84 (m, 2H), 7.47-7.39 (m, 3H), 7.21 (s,1H), 3.88 (s, 6H). ESI-MS m/z calcd for C₁₉H₁₅F₄NO₆ 429.08, found 316.3[M + H]⁺ Chemical Formula: C₁₉H₁₅F₄NO₆ Exact Mass: 429.08 MolecularWeight: 429.32 Compound 13

¹H-NMR (400 MHz, DMSO): δ 10.26 (br, 1H); 7.55 (s, 1H), 7.37 (s, 1H),3.86 (s, 3H), 3.84 (s, 3H), 1.48 (s, 9H). ESI-MS m/z calcd forC₁₇H₂₀F₃NO₆ 391.12, found 278.3 [M + H]⁺ Chemical Formula: C₁₇H₂₀F₃NO₆Exact Mass: 391.12 Molecular Weight: 391.34 Compound 14

¹H-NMR (300 MHz, DMSO): δ 7.41 (s, 1H), 7.33-7.30 (d, 1H), 7.01 (s, 1H),6.73 (s, 1H), 6.67-6.64 (d, 1H), 3.85-3.84 (s, 6H), 3.82 (s, 3H), 3.76(s, 3H). ESI-MS m/z calcd for C₂₁H₂₀F₃NO₈ 471.11, found 358.3 [M + H]⁺Chemical Formula: C₂₁H₂₀F₃NO₈ Exact Mass: 471.11 Molecular Weight:471.39 Compound 15

¹H-NMR (300 MHz, DMSO): δ 11.43 (s, 1H), 7.97 (br, 1H), 7.44 (s, 1H),7.41-7.30 (m, 4H), 7.02 (s, 1H), 3.86 (s, 3H), 3.82 (s, 3H), 2.22 (s,3H). ESI-MS m/z calcd for C₂₀H₁₈F₃NO₆ 425.11, found 312.3 [M + H]⁺.Chemical Formula: C₂₀H₁₈F₃NO₆ Exact Mass: 425.11 Molecular Weight:425.36 Compound 16

¹H-NMR (400 MHz, DMSO): δ 8.02-7.99 (d, 1H), 7.94-7.74 (m, 2H),7.62-7.60 (d, 1H), 7.45 (s, 1H), 6.94 (s, 1H), 3.94 (s, 3H), 3.86 (s,3H). ESI-MS m/z calcd for C₂₀H₁₅F₆NO₆ 479.08, found 366.3 [M + H]⁺Chemical Formula: C₂₀H₁₅F₆NO₆ Exact Mass: 479.08 Molecular Weight:479.33 Compound 17

¹H-NMR (400 MHz, DMSO): δ 7.61-7.53 (m, 2H), 7.42 (s, 1H), 7.40-7.33 (m,2H), 6.99 (s, 1H), 3.84 (s, 3H), 3.82 (s, 3H). ESI-MS m/z calcd forC₁₉H₁₅F₄NO₆ 429.08, found 316.3 [M + H]⁺ Chemical Formula: C₁₉H₁₅F₄NO₆Exact Mass: 429.08 Molecular Weight: 429.32 Compound 18

¹H-NMR (400 MHz, DMSO): δ 7.49-7.41 (m, 2H), 7.31 (s, 1H), 7.17-7.04 (m,2H), 3.90- 3.87 (m, 9H), 3.76 (s, 3H). ESI-MS m/z calcd for C₂₁H₂₀F₃NO₈471.11, found 358.3 [M + H]⁺ Chemical Formula: C₂₁H₂₀F₃NO₈ Exact Mass:471.11 Molecular Weight: 471.39 Compound 19

¹H-NMR (300 MHz, DMSO): δ 7.69-7.65 (m, 2H), 7.59-7.54 (m, 2H), 7.44 (s,1H), 7.00 (s, 1H), 3.86 (s, 3H), 3.84 (s, 3H). ESI-MS m/z calcd forC₂₀H₁₅F₆NO₇ 495.08, found 382.3 [M + H]⁺ Chemical Formula: C₂₀H₁₅F₆NO₇Exact Mass: 495.08 Molecular Weight: 495.33 Compound 20

¹H-NMR (300 MHz, DMSO): δ 7.65-7.57 (m, 1H), 7.50-7.42 (m, 2H),7.20-7.14 (m, 1H), 7.10- 7.00 (m, 2H), 4.40-4.33 (q, 2H), 3.86 (s, 3H),3.83 (s, 3H), 1.23-1.19 (t, 3H). ESI-MS m/z calcd for C₂₁H₂₀F₃NO₇455.12, found 342.3 [M + H]⁺ Chemical Formula: C₂₁H₂₀F₃NO₇ Exact Mass:455.12 Molecular Weight: 455.39 Compound 21

¹H-NMR (400 MHz, DMSO): δ 7.47-7.40 (m, 3H), 7.18-7.16 (d, 1H),7.08-7.04 (m, 1H), 7.00 (s, 1H), 3.97-3.94 (t, 2H), 3.85 (s, 3H), 3.81(s, 3H), 1.62-1.56 (m, 2H), 0.84-0.80 (t, 3H). ESI-MS m/z calcd forC₂₂H₂₂F₃NO₇ 469.13, found 356.3 [M + H]⁺ Chemical Formula: C₂₂H₂₂F₃NO₇Exact Mass: 469.13 Molecular Weight: 469.41 Compound 22

¹H-NMR (400 MHz, DMSO): δ 12.11 (br, 1H), 8.13 (dd, 1H), 7.91-7.82 (m,2H), 7.62 (dd, 1H), 7.48 (s, 1H), 6.98 (s, 1H), 3.88 (s, 3H), 3.83 (s,3H), 3.17 (s, 3H). ESI-MS m/z calcd for C₂₀H₁₈F₃NO₈S 489.07, found 376.3[M + H]⁺ Chemical Formula: C₂₀H₁₈F₃NO₈S Exact Mass: 489.07 MolecularWeight: 489.42 Compound 23

¹H-NMR (300 MHz, DMSO): δ 7.85 (s, 1H), 7.60 (s, 2H), 7.44 (s, 1H), 6.95(s, 1H), 3.90 (s, 3H), 3.84 (s, 3H). ESI-MS m/z calcd for C₁₉H₁₄Cl₂F₃NO₆479.02, found 366.2 [M + H]⁺ Chemical Formula: C₁₉H₁₄Cl₂F₃NO₆ ExactMass: 479.02 Molecular Weight: 480.22 Compound 24

¹H-NMR (400 MHz, DMSO): δ 7.59-7.57 (d, 1H), 7.54-7.49 (m, 1H),7.46-7.41 (m, 2H), 7.38- 7.34 (m, 1H), 7.22-7.19 (d, 1H), 7.11-7.08 (m,1H), 3.77 (s, 3H). ESI-MS m/z calcd for C₁₈H₁₃BrF₃NO₅ 458.99, found346.2 [M + H]+ Chemical Formula: C₁₈H₁₃BrF₃NO₅ Exact Mass: 458.99Molecular Weight: 460.20 Compound 25

¹H-NMR (300 MHz, DMSO): δ 7.52-7.47 (m, 1H), 7.41-7.39 (m, 2H),7.21-7.18 (d, 1H), 7.14- 7.06 (m, 1H), 6.98 (s, 1H), 6.13 (s, 2H), 3.77(s, 3H). ESI-MS m/z calcd for C₁₉H₁₄F₃NO₇ 425.07, found 312.2 [M + H]⁺Chemical Formula: C₁₉H₁₄F₃NO₇ Exact Mass: 425.07 Molecular Weight:425.32 Compound 26

¹H-NMR (300 MHz, CD3OD): δ 7.72 (s, 1H), 7.57 (s, 1H), 7.16 (s, 1H),6.90 (s, 1H), 4.03 (s, 3H), 4.02-4.00 (m, 6H), 3.92 (s, 3H). ESI-MS m/zcalcd for C₂₁H₁₉BrF₃NO₈ 549.02, found 436.2 [M + H] Chemical Formula:C₂₁H₁₉BrF₃NO₈ Exact Mass: 549.02 Molecular Weight: 550.28 Compound 27

¹H-NMR (400 MHz, DMSO): δ 11.78 (br, 1H), 8.16 (d, 1H), 7.92 (d, 1H),7.89-7.74 (m, 2H), 7.44 (s, 1H), 7.00 (s, 1H), 3.86 (s, 6H). ESI-MS m/zcalcd for C₁₉H₁₅F₃N₂O₈ 456.08, found 343.3 [M + H]⁺ Chemical Formula:C₁₉H₁₅F₃N₂O₈ Exact Mass: 456.08 Molecular Weight: 456.33 Compound 29

¹H-NMR (400 MHz, DMSO): δ 11.29 (br, 1H), 7.43 (s, 1H), 7.21 (d, 2H),7.09 (s, 1H), 6.82 (d, 1H), 6.71 (m, 2H), 4.96 (br, 2H), 3.92 (s, 3H),3.85 (s, 3H). ESI-MS m/z calcd for C₁₇H₁₆N₂O₄ 312.11, found 313.3 [M +H]⁺ Chemical Formula: C₁₇H₁₆N₂O₄ Exact Mass: 312.11 Molecular Weight:312.33 Compound 30

¹H-NMR (300 MHz, DMSO): δ 8.55 (s, 1H), 8.30-8.27 (d, 1H), 8.03-8.00 (d,2H), 7.85- 7.79 (m, 3H), 7.16-7.10 (m, 4H), 5.76 (s, 2H), 3.90 (s, 3H),3.87 (s, 3H). ESI-MS m/z calcd for C₂₈H₂₂F₃NO₉ 573.12, found 460.4 [M +H]⁺ Chemical Formula: C₂₈H₂₂F₃NO₉ Exact Mass: 573.12 Molecular Weight:573.48 Compound 32

¹H-NMR (300 MHz, DMSO): δ 8.41 (s, 1H), 8.23-8.20 (d, 1H), 7.81-7.78 (d,2H), 7.75- 7.72 (d, 1H), 7.15-7.12 (d, 2H), 3.84 (s, 3H). ESI-MS m/zcalcd for C₁₇H₁₃NO₅ 311.08, found 312.3 [M + H]⁺ Chemical Formula:C₁₇H₁₃NO₅ Exact Mass: 311.08 Molecular Weight: 311.29 Compound 33

¹H-NMR (400 MHz, CDCl₃): δ 7.94-7.93 (d, 1H), 7.54-7.50 (m, 1H),7.07-7.03 (m, 1H), 6.91- 6.89 (d, 1H), 6.87-6.81 (m, 3H), 3.82 (s, 3H),3.78 (s, 3H). ESI-MS m/z calcd for C₁₈H₁₅NO₅ 325.10, found 348.3 [M +Na] Chemical Formula: C₁₈H₁₅NO₅ Exact Mass: 325.10 Molecular Weight:325.32 Compound 34

¹H-NMR (400 MHz, CDCl₃): δ 7.63-7.60 (dd, 1H), 7.52 (s, 1H), 7.49-7.45(m, 1H), 7.44- 7.36 (m, 2H), 6.93 (s, 1H), 4.08-4.06 (d, 2H), 4.00 (s,3H), 3.96 (s, 3H). ESI-MS m/z calcd for C₁₇H₁₅ClN₂O₃ 330.08, found 331.1[M + H]⁺ Chemical Formula: C₁₇H₁₅ClN₂O₃ Exact Mass: 330.08 MolecularWeight: 330.77 Compound 35

¹H-NMR (400 MHz, CD3OD): δ 7.67-7.65 (d, 1H), 7.61-7.52 (m, 4H), 7.02(s, 1H), 3.95 (s, 3H), 3.92 (s, 3H). ESI-MS m/z calcd for C₁₇H₁₅ClN₂O₃330.08, found 331.1 [M + H]⁺ Chemical Formula: C₁₇H₁₅ClN₂O₃ Exact Mass:330.08 Molecular Weight: 330.77 Compound 36

¹H-NMR (300 MHz, DMSO): δ 7.70-7.64 (d, 2H), 7.58-7.46 (m, 3H), 7.41 (s,1H), 7.10 (s, 1H), 4.16 (br, 2H), 3.96 (s, 3H), 3.82 (s, 3H). ESI-MS m/zcalcd for C₁₇H₁₆Cl₂N₂O₃ 296.12, found 297.3 [M + H]⁺ Chemical Formula:C₁₇H₁₆N₂O₃ Exact Mass: 296.12 Molecular Weight: 296.33 Compound 37

¹H-NMR (300 MHz, DMSO): δ 8.40-8.36 (m, 2H), 7.58-7.56 (d, 1H),7.53-7.48 (dd, 1H), 7.33 (s, 1H), 7.18-7.15 (d, 1H), 7.07-7.02 (dd, 1H),6.90 (s, 1H), 3.94 (s, 3H), 3.84-3.81 (m, 6H). ESI-MS m/z calcd forC₁₈H₁₈N₂O₄ 326.13, found 327.3 [M + H]⁺ Chemical Formula: C₁₈H₁₈N₂O₄Exact Mass: 326.13 Molecular Weight: 326.35 Compound 38

¹H-NMR (300 MHz, DMSO): δ 8.31-8.28 (d, 1H), 8.09-7.78 (m, 1H),7.76-7.69 (m, 2H), 7.40- 7.35 (d, 2H), 7.12-7.09 (d, 2H), 3.84 (s, 3H),3.55 (s, 3H). ESI-MS m/z calcd for C₁₇H₁₅NO₃ 281.11, found 282.3 [M +H]⁺ Chemical Formula: C₁₇H₁₅NO₃ Exact Mass: 281.1052 Molecular Weight:281.3110 Compound 39

¹H-NMR (300 MHz, CDCl₃): δ 7.97-7.94 (d, 1H), 7.56-7.33 (m, 4H),7.15-7.04 (m, 6H), 3.94 (s, 3H), 3.07 (s, 6H). ESI-MS m/z calcd forC₂₄H₂₂N₂O₃ 386.16, found 387.4 [M + H]⁺ Chemical Formula: C₂₄H₂₂N₂O₃Exact Mass: 386.16 Molecular Weight: 386.45 Compound 40

¹H-NMR (300 MHz, DMSO): δ 7.98-7.96 (d, 1H), 7.90-7.79 (m, 2H),7.69-7.67 (d, 1H), 7.56 (s, 1H), 7.09 (s, 1H), 3.91 (s, 3H), 3.80 (s,3H). ¹⁹F-NMR (300 MHz, DMSO): δ −58.54. ESI-MS m/z calcd for C₁₈H₁₄F₃NO₄365.09, found 366.3 [M + H]⁺ Chemical Formula: C₁₈H₁₄F₃NO₄ Exact Mass:365.09 Molecular Weight: 365.31 Compound 41

¹H-NMR (300 MHz, DMSO): δ 8.00-7.98 (d, 1H), 7.92-7.81 (m, 2H),7.73-7.71 (d, 1H), 7.68 (s, 1H), 7.23 (s, 1H), 3.94 (s, 3H), 3.90 (s,3H). ¹⁹F-NMR (300 MHz, DMSO): δ −58.49. ESI-MS m/z calcd forC₁₈H₁₅ClF₃NO₄ 401.06, found 366.3 [M + H]⁺ Chemical Formula:C₁₈H₁₅ClF₃NO₄ Exact Mass: 401.06 Molecular Weight: 401.77 Compound 42

¹H-NMR (400 MHz, DMSO): δ 7.94-7.92 (d, 1H), 7.85-7.78 (m, 2H),7.76-7.60 (d, 1H), 7.44 (s, 1H), 6.99 (s, 1H), 3.86 (s, 3H), 3.83 (s,3H). ¹⁹F-NMR (400 MHz, DMSO): δ −58.63. ESI-MS m/z calcd for C₂₀H₁₈F₃NO₆425.11, found 366.3 [M + H]⁺ Chemical Formula: C₂₀H₁₈F₃NO₆ Exact Mass:425.11 Molecular Weight: 425.36 Compound 43

¹H-NMR (400 MHz, DMSO): δ 7.98-7.96 (d, 1H), 7.89-7.79 (m, 2H),7.69-7.67 (d, 1H), 7.56 (s, 1H), 7.09 (s, 1H), 3.93 (s, 3H), 3.87 (s,3H). ¹⁹F-NMR (400 MHz, DMSO): δ −58.54. ESI-MS m/z calcd forC₁₉H₁₈F₃NO₇S 461.08, found 366.3 [M + H]⁺ Chemical Formula: C₁₉H₁₈F₃NO₇SExact Mass: 461.08 Molecular Weight: 461.41 Compound 44

¹H-NMR (400 MHz, DMSO): δ 7.88-7.86 (d, 2H), 7.61-7.59 (d, 2H), 7.41 (s,1H), 7.13 (s, 1H), 3.85 (s, 6H). ESI-MS m/z calcd for C₁₇H₁₄ClNO₄331.06, found 332.2 [M + H]⁺ Chemical Formula: C₁₇H₁₄ClNO₄ Exact Mass:331.06 Molecular Weight: 331.75 Compound 45

¹H-NMR (400 MHz, DMSO): δ 7.64- 7.48 (m, 4H), 7.44 (s, 1H), 6.97 (s,1H), 3.85 (s, 3H), 3.82 (s, 3H). ESI-MS m/z calcd for C₁₇H₁₄ClNO₄331.06, found 332.3 [M + H]⁺ Chemical Formula: C₁₇H₁₄ClNO₄ Exact Mass:331.06 Molecular Weight: 331.75 Compound 46

¹H-NMR (400 MHz, DMSO): δ 7.86-7.82 (m, 2H), 7.43-7.37 (m, 3H), 7.15 (s,1H), 3.85 (s, 6H). ESI-MS m/z calcd for C₁₇H₁₄FNO₄ 315.09, found 316.3[M + H]⁺ Chemical Formula: C₁₇H₁₄FNO₄ Exact Mass: 315.09 MolecularWeight: 315.30 Compound 47

¹H-NMR (300 MHz, DMSO): δ 7.67-7.65 (d, 2H), 7.59-7.54 (dd, 2H), 7.44(s, 1H), 6.99 (s, 1H), 3.86 (s, 3H), 3.84 (s, 3H). ESI-MS m/z calcd forC₁₈H₁₄F₃NO₅ 381.08, found 382.3 [M + H]⁺ Chemical Formula: C₁₈H₁₄F₃NO₅Exact Mass: 381.08 Molecular Weight: 381.31 Compound 48

¹H-NMR (400 MHz, DMSO): δ 7.84 (s, 1H), 7.60 (s, 2H), 7.44 (s, 1H), 6.95(s, 1H), 3.85 (s, 6H). ESI-MS m/z calcd for C₁₇H₁₃Cl₂NO₄ 365.02, found366.0 [M + H]⁺ Chemical Formula: C₁₇H₁₃Cl₂NO₄ Exact Mass: 365.02Molecular Weight: 366.19 Compound 49

¹H-NMR (300 MHz, DMSO): δ 8.15 (dd, 1H), 7.91-7.78 (m, 2H), 7.59 (dd,1H), 7.42 (s, 1H), 6.96 (s, 1H), 3.84 (s, 3H), 3.82 (s, 3H), 3.18 (s,3H). ESI-MS m/z calcd for C₁₈H₁₇NO₆S 375.08, found 376.1 [M + H]⁺Chemical Formula: C₁₈H₁₇NO₆S Exact Mass: 375.08 Molecular Weight: 375.40Compound 50

¹H-NMR (300 MHz, DMSO): δ 7.58-7.55 (d, 1H), 7.51 (s, 1H), 7.47-7.44 (d,1H), 7.25- 7.22 (d, 1H), 7.16-7.09 (m, 2H), 3.90 (s, 3H), 3.86 (s, 3H),3.77 (s, 3H). ESI-MS m/z calcd for C₁₈H₁₈ClNO₅ 363.09, found 328.3 [M +H]⁺ Chemical Formula: C₁₈H₁₈ClNO₅ Exact Mass: 363.09 Molecular Weight:363.79 Compound 51

¹H-NMR (300 MHz, DMSO): δ 7.74-7.70 (m, 2H), 7.63-7.54 (m, 3H),7.16-7.13 (m, 1H), 3.91- 3.88 (m, 6H). ESI-MS m/z calcd forC₁₈H₁₅ClF₃NO₅ 417.06, found 382.3 [M + H]⁺ Chemical Formula:C₁₈H₁₅ClF₃NO₅ Exact Mass: 417.0591 Molecular Weight: 417.7652 Compound52

Biological Examples of the Invention

Another aspect of the invention relates to methods of inhibiting NADPHoxidase (NOX) in cell, comprising the step of administering to the cellan effective amount of the compound or the pharmaceutical compositionaccording to embodiments of the present application.

Within the context according to the present application, samples withNOX activity include enzymes; tissue or cell cultures; biologicalsamples such as biological material samples (blood, serum, urine,cerebrospinal fluid, tears, sputum, saliva, tissue samples, and thelike); laboratory samples; biological samples such as extracts of cells,particularly recombinant cells expressing NOX; and the like. Samples canbe contained in any medium including water and organic solvent\watermixtures.

If desired, the NOX inhibiting activity of a compound of the presentdisclosure after use of the compound or the composition can be observedby any method including direct and indirect methods of detecting suchactivity. Quantitative, qualitative, and semiquantitative methods ofdetermining such activity are all contemplated. Typically one of themethods described below are applied, however, any other method such asobservation of the physiological properties of a living organism arealso applicable.

The NOX inhibiting activity of a compound or a composition according tothe present application can be measured using standard screeningprotocols that are known. For example, the NOX inhibiting activity of acompound or a composition can be measured using the following generalprotocols.

NOX1 HT29 Cell-Based Assay:

HT29 cells were cultured in T75 flask (Corning) and when 70-80%confluence was reached, cells were trypsinized, harvested in HBSS andcounted. 1-2×10⁵ cells were dispensed into individual wells in 70 μlfinal volume (96-well plates, white Corning). Cells were treated for30-60 mins at 37° C. with 10 μl of GKT137831, DMSO and test compoundsfrom their respective assay plates. This will correspond to a finalconcentration of 100 nM, 1000 nM GKT137831 or test compounds, and 0.1%DMSO. 20 μL of a mixture containing 100 μM luminol plus 0.4 units of HRP(final concentration) was added. Luminescence was quantified using aSpectraMax i3 Multiplate reader (Molecular Devices, Sunnyvale, Calif.,USA). As designed, compounds that inhibit NOX1 activity will reducecellular ROS production, leading to reduced probe-ROS interactions andreduced well luminescence. Compounds were tested in duplicate at aconcentration of 100 nM and 1000 nM. DMSO and GKT137831 wells wereapplied as negative and positive controls, and the activities measuredwere set to 0% and 100%, respectively.

NOX2 HL60 Cell-Based Assay:

HL-60 cells, rich in NOX2 expression, was stimulated to undergooxidative burst by phorbol myristate acetate (PMA). The ROS producedduring this process was measured using the O₂ ⁻-specificchemiluminescent probe L012. L012 was added to DMSO-differentiated HL-60cell suspensions to a final concentration of 100 μM. Then, 80 μLaliquots of cell suspension (2.5×10⁵ cells) were transferred toindividual wells of a 96-well plate. Test compounds were directly addedto the wells at the specified concentrations. Following a 30-60 minsincubation at 37° C. in the dark, the oxidative burst was initiated bythe addition of 10 ng/mL PMA, and the chemiluminescence signal wasrecorded every 5 min over a period of 60 mins in a SpectraMax i3Multiplate reader (Molecular Devices, Sunnyvale, Calif., USA).

NOX4 HEK293 Overexpression Cell-Based Assay:

Extracellular H₂O₂ measurement using Amplex™ Red HydrogenPeroxide/Peroxidase Assay Kit (Catalog number: A22188; Invitrogen,Carlsbad, Calif., USA) was used to detect extracellular H₂O₂ release byHEK293 cells stably overexpressing NOX4. NOX4 clonal cells in log-phasegrowth were trypsinized, washed, and dispersed thoroughly. Followingtrypsinization, the cells (3×10⁴) were resuspended in 100 μL of 1×Krebs-Ringer phosphate glucose (KRPG) buffer containing variousconcentrations of GKT137831 or test compounds and incubated at 37° C.for 30 min. Cells were mixed with 100 μL of Amplex Red reagent solutioncontaining 50 μM Amplex Red and 0.1 units/mL of HRP in KRPG buffer; eachassay condition was evaluated induplicate. The samples were incubated at37° C. for 90 mins in the dark, and then centrifuged at 2,500 g for 5mins. The supernatants were loaded to a 96 well plate, and then measuredusing a SpectraMax i3 Multiplate reader (Molecular Devices, Sunnyvale,Calif., USA) at 530 nm excitation and 590 nm emission respectively. DMSOand GKT137831 wells were applied as negative and positive controls, andthe activities measured were set to 0 and 100, respectively.

Representative examples of the NOX inhibiting activity of the compoundsof Formula I are shown in the Table below. Wherein, A indicates that thepercent of inhibition of NOX by the compounds is larger than or equalsto 80%; B indicates that the percent of inhibition of NOX by thecompounds ranges from 50% o to 79% o; C indicates that the percent ofinhibition of NOX by the compounds ranges from 2000 to 49%; and Dindicates that the percent of inhibition of NOX by the compounds issmaller than or equals to 19% o.

Inhibition of NOX Activity by the Compounds

% Inhibition % Inhibition % Inhibition Compound # NOX1 NOX2 NOX4 (nM)100 1000 100 1000 100 1000 2 A A D B C A 3 C A C A D B 4 B A D B D A 5 AA D A C A 6 A A D D C B 7 A A D B B A 8 C A D D D B 45 D A D D C B 9 A AD A B A 46 A A D A B B 10 A A D C C A 11 D B D D C A 12 D A D D D A 13 AA D A C A 47 A A D A C A 14 D A D D D D 15 A A C A C A 32 D B C C D C 16A A C A B B 17 A A C B B B 33 A A D B D B 39 D A D D D B 18 A A C B C B19 A A D B D B 20 A A B A C B 21 A A D B C A 22 A A C A C A 29 A A C A BB 23 A A B A B B 34 D D D D D D 51 A A C A D B 24 A A C A C B 25 B A D BD C 30 D A B A D D 37 D A B A D B 26 A A B A C A 52 A A C B C B 27 A A DA C A 42 A A B A B B 43 A A B A B B 44 A A C A B B 40 D A C B D B 48 A AC B C A 41 A A C A B B 38 D D D D D C 35 D D D D D D 49 A A D A C B 50 AA B A B B 36 C A C A D B

It will be apparent to those skilled in the art that variousmodifications and variations can be made to the disclosed embodiments.It is intended that the specification and examples be considered asexemplary only, with a true scope of the disclosure being indicated bythe following claims and their equivalents.

1. A compound of Formula I, or a geometric isomer, enantiomer,diastereomer, racemate, atropisomer, pharmaceutically acceptable salt,prodrug or solvate thereof:

Wherein A is C₆-C₂₀ aryl optionally fused with C₁-C₂₀ heterocyclyl, orC₁-C₂₀ heteroaryl optionally fused with C₁-C₂₀ heterocyclyl, and A isoptionally substituted with one or more C₁-C₁₀ alkyl, C₆-C₂₀ aryl,halogen, OR₁₁, C(O)R₁₁, C(O)OR₁₁, NR₁₁R₁₂, SO₂R₁₁, or SO₂(OR₁₁); B isC₁-C₁₀ alkyl, C₆-C₂₀ aryl, C₁-C₂₀ heteroaryl or C₁-C₂₀ heterocyclyl, andB is optionally substituted with one or more C₁-C₁₀ alkyl, C₆-C₂₀ aryl,C₁-C₂₀ heterocyclyl, halogen, OR₂₁, C(O)R₂₁, C(O)OR₂₁, NR₂₁R₂₂, SO₂R₂₁,or SO₂(OR₂₁), wherein said C₁-C₁₀ alkyl is optionally substituted withone or more halogen; R₁₁, R₁₂, R₂₁ and R₂₂ independently are H, O,C₁-C₁₀ alkyl, or C₆-C₂₀ aryl, wherein said C₁-C₁₀ alkyl is optionallysubstituted with one or more halogen;

is a single bond or double bond; n is 0 or 1; X is N or C; Y is H, OH orNH₂; Z is nil or C(O); R₃ and R₄ independently are nil, H, C₁-C₁₀ alkylor C₆-C₂₀ aryl; and R₅ is nil, H or C₁-C₁₀ alkyl.
 2. The compound ofclaim 1 represented by Formula II,

wherein A is C₆-C₂₀ aryl optionally fused with C₁-C₂₀ heterocyclyl, orC₁-C₂₀ heteroaryl optionally fused with C₁-C₂₀ heterocyclyl, and A isoptionally substituted with one or more C₁-C₁₀ alkyl, C₆-C₂₀ aryl,halogen, OR₁₁, C(O)R₁₁, C(O)OR₁₁, NR₁₁R₁₂, SO₂R₁₁, or SO₂(OR₁₁); B isC₁-C₁₀ alkyl, C₆-C₂₀ aryl, C₁-C₂₀ heteroaryl or C₁-C₂₀ heterocyclyl andis optionally substituted with one or more C₁-C₁₀ alkyl, C₆-C₂₀ aryl,C₁-C₂₀ heterocyclyl, halogen, OR₂₁, C(O)R₂₁, C(O)OR₂₁, NR₂₁R₂₂, SO₂R₂₁,or SO₂(OR₂₁), wherein said C₁-C₁₀ alkyl is optionally substituted withone or more halogen; R₁₁, R₁₂, R₂₁ and R₂₂ independently are H, O,C₁-C₁₀ alkyl, or C₆-C₂₀ aryl, wherein said C₁-C₁₀ alkyl is optionallysubstituted with one or more halogen; Y is OH or NH₂; Z is nil or C(O);and R₅ is nil, H, or C₁-C₁₀ alkyl.
 3. The compound of claim 1represented by Formula III,

Wherein A is C₆-C₂₀ aryl optionally fused with C₁-C₂₀ heterocyclyl, orC₁-C₂₀ heteroaryl optionally fused with C₁-C₂₀ heterocyclyl, and A isoptionally substituted with one or more C₁-C₁₀ alkyl, C₆-C₂₀ aryl,halogen, OR₁₁, C(O)R₁₁, C(O)OR₁₁, NR₁₁R₁₂, SO₂R₁₁, or SO₂(OR₁₁); B isC₁-C₁₀ alkyl, C₆-C₂₀ aryl, C₁-C₂₀ heteroaryl or C₁-C₂₀ heterocyclyl, andB is optionally substituted with one or more C₁-C₁₀ alkyl, C₆-C₂₀ aryl,C₁-C₂₀ heterocyclyl, halogen, OR₂₁, C(O)R₂₁, C(O)OR₂₁, NR₂₁R₂₂, SO₂R₂₁,or SO₂(OR₂₁), wherein said C₁-C₁₀ alkyl is optionally substituted withone or more halogen; R₁₁, R₁₂, R₂₁ and R₂₂ independently are H, O,C₁-C₁₀ alkyl, or C₆-C₂₀ aryl, wherein said C₁-C₁₀ alkyl is optionallysubstituted with one or more halogen; Z is nil or C(O); and each R₃ andR₄ independently is nil, H, C₁-C₁₀ alkyl or C₆-C₂₀ aryl.
 4. The compoundof claim 1, wherein A is substituted or unsubstituted C₆-C₂₀ aryl orC₆-C₂₀ aryl fused with C₁-C₂₀ heterocyclyl.
 5. The compound of claim 4,wherein A is phenyl group substituted with one or more methoxy group,(dimethylamide) phenyl group, Br, OR₁₁, or C(O)OR₁₁, wherein Ru is H, O,C₁-C₁₀ alkyl, or C₆-C₂₀ aryl, wherein said C₁-C₁₀ alkyl is optionallysubstituted with one or more halogen.
 6. The compound of claim 1,wherein A is substituted or unsubstituted C₁-C₂₀ heteroaryl having atleast one heteroatom selected from S, N, and O.
 7. The compound of claim1, wherein A is substituted or unsubstituted C₆-C₂₀ aryl fused withC₁-C₂₀ heterocyclyl having at least one heteroatom selected from S, N,and O.
 8. The compound of claim 1, wherein B is substituted orunsubstituted C₁-C₁₀ alkyl.
 9. The compound of claim 1, wherein B issubstituted or unsubstituted C₆-C₂₀ aryl.
 10. The compound of claim 12,wherein the B is a phenyl group substituted with one or more methyl,CF₃, halogen, pyrrolidine, OR₂₁, C(O)R₂₁, NR₂₁R₂₂, or SO₂R₂₁.
 11. Thecompound of claim 13, wherein R₂₁ is H, O, methyl, ethyl, n-propyl, orCF₃; and R₂₂ is H or O.
 12. The compound of claim 1, wherein B issubstituted or unsubstituted C₁-C₂₀ heteroaryl.
 13. The compound ofclaim 1, wherein n is
 0. 14. The compound of claim 1, wherein X is C.15. The compound of claim 18, wherein Y is OH or NH₂.
 16. The compoundof claim 1, wherein Z is nil.
 17. The compound of claim 1, wherein R₅ isnil, H or methyl.
 18. The compound of claim 1, wherein the compound isselected from a group consisting of:


19. A pharmaceutical composition comprising a compound of claim 1 and atleast a pharmaceutically acceptable excipient or carrier.
 20. A methodof inhibiting NADPH oxidase (NOX), comprising: administering to a cellan effective amount of the compound as recited in claim 1.